Previous Results:

• The final result of the phage precipitation was a pellet in the bottom of the 50 mL tube. This was the condensed phage that will be used for DNA extraction

Objective:

• To complete DNA extraction without contamination and obtain enough DNA to continue analyzing it
• Have enough time to complete the nanodrop process and have positive results

Procedure:

1. The 50 mL tube containing the phage pellet was brought to room temp
2. 0.5 ml sterile water was added to tube and mixed to resuspend the pellet
3. 2 ml of DNA Clean-Up Resin at 37 degrees Celsius was added and mixed
4. Pellet was separated equally into 2 microcentrifuge tubes and spun at 12.5 x 1000 g for 3 minutes
5. Supernatant was removed from the microcentrifuge tubes without disturbing the cloudy precipitant (DNA) and add 1 ml 80% Isopropanol to the tubes each
6. Step 5 was repeated 2 times, then once more but without removing the alcohol
7. Solution was then transferred to two columns and the vacuum was used to filter the liquid
8. Columns were centrifuged at 12,000 g for 5 min
9. 100 microliters of Elution Buffer at 80 degrees Celsius was added to each column. Left to sit for 1 minute
10. Columns were centrifuged at 12,000 g for 1 minute
11. Two tubes of DNA sample were combined into 1 microcentrifuge tube and labeled. Phage was named “Pippa”
12. The nanodrop machine was then used by adding a drop of Elution Buffer to the machine to prepare for DNA drop
13. DNA drop was added and the machine provided results

Results:

• The results of the nanodrop showed the sample of DNA was not concentrated enough and had a nucleic acid reading of 43.1 ng/uL

Next Steps:

• A PCR will be dome on the DNA sample and a gel will be completed